The long-term objective of this project is to develop a new technology that will enable rapid, single-molecule detection and identification of DNA sequences present in a biological sample. The current effort will focus on detecting nucleic acid molecules labeled with varying sizes of nanoparticles by recording changes in ionic current through a small, nanometer-scale channel in a """"""""nanopipette."""""""" Once this detection technology has been demonstrated, the labeled oligonucleotides can be hybridized to a test sample, the unhybridized labeled molecules removed, and the remaining labeled DNA molecules can be rapidly detected on a single-molecule basis through the nanopipette. This will result in an ultra-sensitive, rapid genotyping technology that can be used for point-of-care diagnostics. The diagnostics can include the detection of pathogens or the determination of a human genotype in a clinical sample. This nanopipette DNA detection technology will also pave the way for second-generation devices, which allow higher resolution detection and could be used for rapid, single-molecule DNA sequencing, eventually realizing the possibility of sequencing an entire human genome in a matter of seconds. In this effort, Stanford will develop and demonstrate this nucleic acid detection technology with the following Specific Aims: -Nanopipette fabrication and characterization -Labeling DNA with nanoparticles -Measurement of labeled DNA

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HG003448-02
Application #
6942314
Study Section
Special Emphasis Panel (ZRG1-ISD (01))
Program Officer
Schloss, Jeffery
Project Start
2004-09-01
Project End
2006-05-31
Budget Start
2005-06-01
Budget End
2006-05-31
Support Year
2
Fiscal Year
2005
Total Cost
$195,313
Indirect Cost
Name
Stanford University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Umehara, Senkei; Pourmand, Nader; Webb, Chris D et al. (2006) Current rectification with poly-l-lysine-coated quartz nanopipettes. Nano Lett 6:2486-92
Karhanek, Miloslav; Kemp, Jennifer T; Pourmand, Nader et al. (2005) Single DNA molecule detection using nanopipettes and nanoparticles. Nano Lett 5:403-7